- Description: Visual
- Average Weight: Check weight of 20 tablets at random and calculate the average weight by formula.
Average weight (mg) = wt of 20 tablets (gm) x 1000
20
3.0) Uniformity of weight:
Weigh 20 tablets selected at random and calculate the average weight. Not more than two of the individual weights deviate from the average weight by more than the percentage shown in table.
Weigh the tablets individually and calculate the percentage of deviation for each tablet by using formula.
Deviation (%) = (experimental weight – theoretical weight) x 100
Theoretical weight
| Average weight of tablets | Percentage deviation |
| 250mg or More | ±5% |
4.0) Identification Test:
Aceclofenac,Paracetamol and Serratiopeptidase:
In the assay, the principal peak in the chromatogram obtained with the test solution corresponds to the principal peak in the chromatogram obtained with the reference solution.
5.0) Hardness:
The standard method used for tablet hardness testing is compression testing. The tablet is placed between two jaws that crush the tablet. The machine measures the force applied to the tablet and detects when it fractures.
6.0) Disintegration Test:
Unless otherwise stated in the individual monograph, introduce one tablet into each tube and add a disc to each tube. The assembly is suspended in the liquid medium in a suitable vessel, preferably a 1-litre beaker. The volume of liquid is such that the wire mesh at its highest point is at least 15mm below the surface of the liquid, and at its lower point is at least 25mm above the bottom of the beaker. At no time should the top of the basket-rack assembly become submerged. There is a thermostatic arrangement for heating the liquid and maintaining the temperature at 37±2°.
If 1 or 2 tablets fail to disintegrate, repeat the test on 12 additional tablets; not less than 16 of the total of 18 tablets tested disintegrate.
If the tablets adhere to the disc and the preparation under examination fails to comply, repeat the test omitting the disc. The preparation complies with the test if all the tablets in the repeat test disintegrate.
7.0) Dimension of Tablet:
Length, breadth and thickness are determined by vernier calliper in mm.
8.0) Leak test:
The apparatus is used to test for the integrity of packed strips, blisters and Alu-Alu Blister pack containing tablets. Ensure apparatus bath is filled with purified water up to mark indicated and add 0.5% crystal violet solution in water. Samples are placed into the desiccators and the lid is placed in position. The pump starts to produce a vacuum 15inHg inside the desiccators and the vacuum is held for 1 minute. The sample remains at the required vacuum level for given time interval buzzer will sound after time is over and will cut off the vacuum pump. As the package is immersed in a colored dye solution the venting of the desiccators will allow any holes to be penetrated by the dye and the contents of the flexible packaging will also be stained with the same coloring material.
Examine all the strips for any leakage by opening the pockets manually. If anyone pocket shows evidence of leakage, reject the sample, stop the Blister / Strip machine and immediately take corrective action.
9.0) Assay:
For: Aceclofenac and Paracetamol:
Determined by liquid chromatography:
Standard solution: Weigh accurately about 162.5 mg of Paracetamol reference standard, and 50 mg of Aceclofenac reference standard in 100ml volumetric flask, dissolve in and dilute to the mark with mobile phase. Shake well and further dilute 2 ml to 25 ml with mobile phase. Shake well and filter through 0.45µm nylon filter.
Test Solution: Weigh accurately about equivalent to 162.5 mg of Paracetamol from crushed powder of 20 tablets in 100 ml volumetric flask, add 50 ml of mobile phase, sonicate for 5 minutes, cool to room temperature and dilute to the mark with mobile phase, filter. Shake well and further dilute 2 ml to 25 ml with mobile phase, shake well and filter through 0.45µm nylon filter.
Procedure: Inject separately standard and test preparation. Inject five replicate injection of working standard and two replicate injection of test preparation. RSD of replicate injection should not more than 2%. Calculate the assay of Paracetamol and Aceclofenac by comparison to peak area of standard preparation.
Chromatographic system:
- A stainless steel column (150cm X4.6mm,) C18
- Mobile phase: Methanol :Water and G.A.A. (80: 20) and (0.01ml); degas
- Flow rate: 1.2ml per minute,
- Spectrophotometer set at 280nm,
- Injection volume: 20µl
Inject the reference solution and test solution.
Calculate the content of Paracetamol and Aceclofenac in tablets.
Formula:
Area of Test STD Wt.(mg) 2 100 25 Potency
—————–X————–X———-X—————–X——X———–X Average weight
Area of STD 100 25 Test Wt. (mg) 2 100
Acceptance criteria: 90.0%-110.0%
For: Serratiopeptidase
Assay. The tyrosine units are dissolved in sodium carbonate solution, which is alkaline in nature. The Folin s ciocalteau reagent helps in colour development where the tyrosine units
bind to the copper molecule in the reagent and causes the reduction of phosphomolybdate which is present in the reagent. There is formation of tyrosine-copper molybdate coloured complex. The intensity ofcolour depends upon the tyrosine units present which is read at 660 nm.
Disodium tetraborate buffer. Dissolve 19.0 gm of disodium tetraborate in 900 ml of water, adjusted to pH 9.0 with 1 M hydrochloric acid and dilute to 1000 ml with water. Casein hammerstein substrate. Dissolve 1.2 gm of Casein Hammerstein to 100 ml ofthe sodium tetraborate buffer. Allow
the casein to dissolve and form homogeneous solution. Boil the solution in boiling water-bath for 2 minutes. After removing from boiling water-bath, cool the casein substrate immediately
in ice cold water. Filter this solution through cotton plug to get clear solution and dilute to 200 ml with water.
NOTE-Always add casein to the buffer while stirring. Do not add buffer to casein as this will cause lumps and not go into solution completely and giving incorrect values. Trichloroacetic acid reagent. Dissolve 18 gm of trichloroacetic Acid, 30 gm of anhydrous sodium acetate and 20 ml ofglacial acetic acid in 1000 ml ofwater. NOTE-TCA should be in the crystalform and not in liquid form as this will effect the quantity weighed and thereby interfere with the complete precipitation of casein. Use
Sodium acetate anhydrous because if, sodium acetate trihydrate should be used, the trihydrate forms gives lowerresults.Diluted Folin ‘s Ciocalteau Reagent. Dilute 1 ml of Fo1in’sCiocalteau reagent with 2 ml ofwater to get 3 ml ofthe reagent.
NOTE-Folin’s Ciocalteau reagent should be ofthe protein estimation grade and not the indicator grade. Folin ‘s reagent . should be diluted to just before addition to the tubes since it is sensitive to light. Do not prepare this solution at the beginning ofthe assay. Tyrosine reference solution. Dissolve 160 mg of tyrosine in 100 ml of 0.2 M hydrochloric acid. Dilute 1 ml ofthis solution to 100 ml with 0.2 Mhydrochloric acid. Use 2 ml ofthis solution for colour development. Trisodium phosphate bufferpH6.8. Weigh 19 g oftrisodium phosphate and 6.4 ml ofhydrochloric acidin 1000 ml ofwater,
adjusted to pH 6.8.
Test solution. Weigh and powder 20 tablets. Disperse a quantity of powder containing about 120 mg of Serratiopeptidase with 100 ml of trisodium phosphate btifferpH6.8. Stir on magnetic stirrer for 1hour. Further dilute 5.0 ml to 25 ml with 5.0percent ammonium sulphate solution, stir for 5 minutes and filter. Dilute 2 ml of the filtrate to 100 ml with sodium tetraborate btiffer. 1 ml of resulting solution is used for analysis. Set the water bath to 37°.
For each sample, keep four test tubes. Mark two test tubes as TEST and two as BLANK. Add 5 ml of Casein Substrate to the tubes marked as TEST and 5 ml. of TCA to the test tube marked as BLANK. Prewarm these tubes for five minutes along with the tubes containing the sample dilutions. Add 1ml of enzyme solution to all these tubes and vertex the tubes for exactly 10seconds. Keep the test tubes in the water-bath at 37° for 20minutes for the reaction. After exactly 20 minutes, add 5 ml of
TCA to tubes marked as TEST and 5 ml of Casein substrate to the tube marked as BLANK. Vertex all the tubes exactly for 30seconds. Keep the tubes in the water-bath at 37° for 30 minutes
for precipitation, filter.
NOTE- For tablets, the settlement may take a longer time than 30 minutes. So filter the solution after the settlement is clearly seen (Vertex in between for better precipitation).
Mark tubes as TEST, BLANK, TYROSINE and TYROSINE
BLANK.
Add the following respectively;
TEST- 2 m1 of the TEST filtrate.
BLANK- 2 ml of BLANK filtrate.
TYROSINE- 2 ml of TYROSINE standard solution.
TYROSINE BLANK- 0.2Mhydrochloric acid.
Add 5 ml of 6 per cent sodium carbonate solution to all tubes. Add 1 ml of diluted Folin’s reagent to all the tubes. Keep the tubes at 37° for 30 minutes for color development.
Measure the absorbance at about 660 nm
A1-A2 1 100 25 100
Serratiopeptidase(mg)=————-X176X———–X————X——X——-X average weight in gm
A3-A4 20 spl. Wt 5 2
Serratiopeptidase Units/Tablet
% of L.A. =——————————————————————–X100
L.A.
Where,
176: Conversion Co-efficient of Tyrosine to serratiopeptidase
20: Reaction time in minutes
DF: Dilution factor
A1: Absorbance due to test preparation
A2: Absorbance due to blank preparation
A3: Absorbance due to standard preparation
A4: Absorbance due to Tyrosine blank
L.A.:20000 Units/tablet
Calculate the % of Serratiopeptidase in the tablets as under:
Activity X100
Serratiopeptidase %: ————————–
20000
Acceptance criteria: N.L.T. 90%
10.0) Abbreviations:
Wt.: Weight
RSD: relative standard deviation
mg: Milligram
`inHg: Inch of Mercury
ml: Millilitre
Std: Standard G.A.A: Glacial acetic acid
